pET-24c(+)

pET-24c(+)

pET24c载体基本信息

出品公司:	Ybscience
别名:	pET24c, pet 24c
质粒类型:	大肠杆菌蛋白表达
表达水平:	高
克隆方法:	多克隆位点，限制性内切酶
载体大小:	5308 bp
5' 测序引物:	T7
5' 测序引物序列:	5'-TAATACGACTCACTATAGGG-3'
载体标签:	N-T7, C-His
载体抗性:	Kanamycin
表达宿主菌:	BL21(DE3) 、BLR(DE3)、BL21(DE3)pLysS
备注:	Same as pET21a,b,c,d(+) but kanamycin resistance; a,b,c,d vary by MCS; The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand.
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

 

pET24c载体质粒图谱和多克隆位点信息

Feature Name	Start	End
T7_terminator	129	1
T7_Terminal_primer	69	87
T7_leader	237	205
Xpress_fwd_primer	238	220
T7_transl_en_RBS	262	246
lacO	307	280
T7_promoter	325	307
tet (300 - 563)	357	620
pBRrevBam_primer	428	409
lacI	703	1794
tet (576 - 851)	1853	2128
ROP	2603	2794
pGEX_3_primer	2810	2788
pBR322_origin	3828	3209
KanR2	3934	4749

ORF	Start	End
ORF frame 3	462	962
ORF frame 1	835	1794
ORF frame 1	3934	4749

Enzyme Name	Cut
XhoI	158
NotI	166
EagI	166
HindIII	173
SalI	179
SacI	190
EcoRI	192
BamHI	198
NheI	229
NdeI	236
XbaI	274
BglII	340
BclI	1076
ApaI	1273
EcoRV	1512
HpaI	1568
FspI	2144
NruI	4022
ClaI	4056
XmaI	4237
SmaI	4239

 

pET24c载体简介

The pET-24a-d(+) vectors carry an N-terminal T7•Tag® sequence plus an optional C-terminal His•Tag® sequence. These vectors differ from pET-21a-d(+) only by their selectable marker (kanamycin vs. ampicillin resistance). Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, singlestranded sequencing should be performed using the T7 terminator primer.

pET24c载体序列

LOCUS       pET-24 c (+)    5308 bp     DNA  circular   SYN
DEFINITION  pET-24 c (+)
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTOAUTHORAD1|Novagen Inc.|
FEATURES             Location/Qualifiers
     source          1..5308
                     /organism="pET-24 c (+)"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    complement(205..237)
                     /label="T7_leader"
     misc_feature    complement(220..238)
                     /label="Xpress_fwd_primer"
     misc_feature    complement(246..262)
                     /label="T7_transl_en_RBS"
     misc_feature    complement(280..307)
                     /label="lacO"
     promoter        complement(307..325)
                     /label="T7_promoter"
     gene            357..620
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(409..428)
                     /label="pBRrevBam_primer"
     CDS             462..962
                     /label="ORF frame 3"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     misc_feature    703..1794
                     /label="lacI"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     CDS             835..1794
                     /label="ORF frame 1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            1853..2128
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    2603..2794
                     /label="ROP"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2788..2810)
                     /label="pGEX_3_primer"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      complement(3209..3828)
                     /label="pBR322_origin"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            3934..4749
                     /label="KanR2"
                     /gene="KanR2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             3934..4749
                     /label="ORF frame 1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
  121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
  181 cgacggagct cgaattcgga tccgacccat ttgctgtcca ccagtcatgc tagccatatg
  241 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
  301 aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg
  361 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
  421 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
  481 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
  541 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
  601 aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt
  661 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
  721 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
  781 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
  841 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
  901 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
  961 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
 1021 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
 1081 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
 1141 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
 1201 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
 1261 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
 1321 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
 1381 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
 1441 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
 1501 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
 1561 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
 1621 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
 1681 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
 1741 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
 1801 cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag
 1861 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
 1921 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat
 1981 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt
 2041 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg
 2101 cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct
 2161 gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc
 2221 gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac
 2281 atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg
 2341 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
 2401 atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat
 2461 ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt
 2521 aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa
 2581 tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg
 2641 ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga
 2701 acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct
 2761 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
 2821 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
 2881 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
 2941 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatatg cggtgtgaaa
 3001 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
 3061 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
 3121 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
 3181 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
 3241 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
 3301 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
 3361 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
 3421 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
 3481 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
 3541 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
 3601 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
 3661 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
 3721 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
 3781 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
 3841 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg
 3901 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
 3961 tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
 4021 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
 4081 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
 4141 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
 4201 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
 4261 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
 4321 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
 4381 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
 4441 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
 4501 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
 4561 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
 4621 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
 4681 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
 4741 tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca
 4801 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gaaattgtaa acgttaatat
 4861 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
 4921 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
 4981 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
 5041 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
 5101 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
 5161 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
 5221 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
 5281 gccgctacag ggcgcgtccc attcgcca
//